Development of two forensically important blowfly species (Chrysomya megacephala and Chrysomya rufifacies) (Diptera: Calliphoridae) at four temperatures in India

The development of the Oriental latrine fly, Chrysomya megacephala (Fabricius), and hairy maggot blowfly, C. rufifacies (Macquart) (Diptera: Calliphoridae), was studied at four different temperatures (22°C, 25°C, 29°C and 31°C) in order to draw correlations between larval age, body length and body dry weight. The mean larval body length increased steadily from a minimum of 1.4 mm for C. megacephala and 1.8 mm for C. rufifacies to a maximum of 17.4 mm for C. megacephala and 15.9 mm for C. rufifacies at different temperatures. Similarly, the mean dry weight increased steadily from a minimum of 0.0007 g for C. megacephala (second instar) and 0.0008 g for C. rufifacies (second instar) to a maximum of 0.0290 g for C. megacephala and 0.0270 g for C. rufifacies at different temperatures. Entomological evidence is often used to estimate the minimum postmortem interval (mPMI) and both of these species are important from a forensic point of view. Graphs of age of larvae vs. body length and age of larvae vs. dry body weight at different temperatures can be used to estimate the larval age of these two species.     

盐酸吗啡对大头金蝇生长发育的影响及其对死者死亡时间推断的意义

用盐酸吗啡注射家兔,处死后用家兔的不同组织饲养大头金蝇 Chrysomya megacephala初孵幼虫,研究吗啡剂量对大头金蝇幼虫生长的影响及其在法医学中推断死者死亡时间方面的应用。结果显示,在28℃下,取食处理组兔肉和肝脏的大头金蝇幼虫的体长和体重均于孵化后28 h开始在不同程度上大于对照组幼虫,这种趋势一直持续到幼虫末期。在实验的剂量范围内（2.67～10.66 mg/kg）,吗啡可促进大头金蝇幼虫的生长。根据大头金蝇幼虫体长和体重推断死者死亡时间时,吗啡的这种影响可使推断值产生的最大偏差达18h。     

二种尸食性蝇类幼虫形态学变化及在法医学上的意义

为了满足实际工作中利用蝇类幼虫日龄推断尸体死后间隔时间的需求,采用形态观察与图像分析相结合的方法对巨尾阿丽蝇Aldrichina grahami(Aldrich)、大头金蝇Chrysomya megacephala(Fabricius)幼虫前气门、后气门、头咽骨进行了研究,筛选适合日龄推断的指标。结果显示:2种尸食性蝇类幼虫前气门、后气门、头咽骨均随着时间的延长发生规律性的变化。其中,咽骨的骨化面积、平均光密度,后气门的平均光密度是最理想的判断幼虫日龄的指标,为法医昆虫学精确推断死后间隔时间提供重要的指导意义。     

Determining the age of adult flesh flies,Boettcherisca peregrina,using pteridine fluorescence

To assess the potential application of pteridine fluorescence in determining the age of adult Boettcherisca peregrina (Diptera: Sarcophagidae) Robineau‐Desvoidy and further for the postmortem interval, the age‐dependent changes of pteridine fluorescence were investigated for the adults maintained at five constant temperatures. From the results, significant linear relationships were found between pteridine fluorescence and the age of the adults maintained at 16, 20, 24, 28 or 32 °C (P < 0.001, r² > 0.85). In addition, the relationships between the rate of pteridine accumulation and temperature were well described using linear equations for adult females and males. Then for each cohort of the flies at the ambient temperature, a calendar was constructed and used to determine the ages of females and males, respectively, in which was recorded in reverse time order the amount of pteridine accumulated per hour by the flies and their expected pteridine level when they emerged at the specified time. A significant linear relationship between estimated ages and chronological ages was observed for female or male adults, with the mean errors of the estimated ages of ±1.82 days for females and ±1.58 days for males. It is suggested that pteridine fluorescence analysis has a potential value in determining the age of adult B. peregrina.     

Amplified fragment length polymorphism confirms reciprocal monophyly in Chrysomya putoria and Chrysomya chloropyga: a correction of reported shared mtDNA haplotypes

Reinvestigation of mitochondrial haplotypes previously reported to be shared between the Afrotropical blowflies Chrysomya putoria Weidemann and Chrysomya chloropyga Weidemann (Diptera: Calliphoridae) revealed an error resulting from the misidentification of specimens. Preliminary amplified fragment length polymorphism (AFLP) analysis of the original and additional individuals again failed to find reciprocal monophyly, leading to a re-examination of the specimens for diagnostic male genitalic characters that were first described following the earlier study. Four of the original study specimens were found to have been misidentified, and definitive analysis of both mtDNA and AFLP genotypes using phylogenetic analysis and genetic assignment showed that each species was indeed reciprocally monophyletic. In addition to correcting the earlier error, this study illustrates how AFLP analysis can be used for efficient and effective specimen identification through both phylogenetic analysis and genetic assignment, and suggests that the latter method has special advantages for identification when no conspecific specimens are represented in the reference database.     

肥须亚麻蝇幼虫头咽骨形态学分析及其法医学意义

为了明确肥须亚麻蝇Parasarcophaga crassipalpis Macguart幼虫头咽骨形态学特征的变化规律,分别在16,20,24,28和32℃下饲养肥须亚麻蝇幼虫并定期取样,显微镜下观察其形态变化,利用图像分析软件测量其口钩和咽骨的面积、平均光密度及骨化面积等形态参数。结果显示:在所测量口钩和咽骨的7项指标中,咽骨的平均光密度及骨化面积是最理想的幼虫日龄的判断指标,对尸体上死后间隔时间的推测具有重要的指导意义;口钩部的4项指标在龄期更叠时有明显变化,仅可进行龄期的鉴别。     

甲虫表皮碳氢化合物研究进展

昆虫表皮碳氢化合物(cuticular hydrocarbons)是正烷烃、不饱和烃以及甲基支链烃的混合物.除了保持水分的基本功能外,还具有多种生物功能.鞘翅目昆虫(俗称甲虫)是多样性最为丰富的昆虫类群,目前对其表皮碳氢化合物研究开展颇多,主要见于化学通讯领域,内容涉及到性信息素、标记信息素、聚集信息素、化学拟态、寄主...     

表皮碳氢化合物分析用于棉铃虫与烟青虫幼虫分类鉴别

本文利用ＧＣ－ＭＳ技术分析了棉铃虫与烟青虫幼虫的表皮碳氢化合物,并与亚洲玉米螟幼虫表皮碳氢化合物进行了比较。结果表明,棉铃虫与烟青虫幼虫表皮碳氢化合物的组分与含量均有显著差异,与玉米螟相比,这二者之间的差异明显小于它们与玉米螟的差异,其表现与三者之间自然的系统发育关系相一致。     

Low temperature episodes in development of blowflies: implications for postmortem interval estimation

Traditionally the calculation of accumulated degree days or hours (ADD or ADH) involves the concept of a minimum threshold temperature below which development ceases. Hence in fluctuating conditions, where temperatures drop below this threshold, there may be periods of time when development is taken to be zero. This has important implications when the calculation of postmortem interval (PMI) is based on the ADD or ADH of larval dipterans. Normal development of larvae of the blowflies Calliphora vicina Robineau-Desvoidy and C. vomitoria L. (Diptera: Calliphoridae) at 20 degrees C was interrupted by cold episodes. The expectation was that total development time would increase by the period at low (therefore no development) temperature but the total ADD or ADH should be the same as non-cold treated cohorts. The results, however, showed that total ADH for both species decreased linearly with increasing temperature with no evidence of any minimum threshold temperature effect. The increased ADH at low temperatures may be due to either continued but reduced development or a delay in development restarting after the cold episode. Use of ADH in PMI estimations has shortcomings particularly during the winter period where low temperatures are involved or where there are sudden summer cold spells during the development period. As blowfly development progresses from egg to pupa such errors will be compounded.     

Seasonality of insect succession on exposed rabbit carrion in Guangzhou, China

Abstract  Succession patterns for insect fauna, spanning four seasons, on rabbit carcasses were studied in Guangzhou China from 2006 to 2008. A total of 49 taxa belonging to three orders and 15 families were collected and identified. Four decomposition stages were recognized: fresh, bloated, decay and dry stage. Taxa diversity was higher in bloated and decay stages than fresh and dry stages. Internal and ambient temperatures were measured. Maximum internal temperature arrived earlier in warmer seasons. The sequence of insect succession in this study follows the same general patterns: the Diptera peaked initially, and the Coleoptera peaked later. Chrysomya megacephala was the dominant species in four seasons. Its time point of different development stages on carrion could be the most important indicators for PMI (postmortem interval) estimation in Guangzhou. Yearly and seasonal differences in activities of insects were compared. Alpha diversity indices were calculated. There were no significant differences in Shannon-Wiener index, evenness ( J ) and dominance ( C ) between seasons and years. From the Jaccard similarity index, the highest similarity was between winter and spring, while the lowest was between summer and winter. Some species occurring exclusively in one season may be important to characterize different seasons. A list of the insect taxa associated with above-ground rabbit carrion during the four seasons was compiled, which can be used for estimating time since death in similar bio-climate areas.     

上海地区最常见嗜尸性蝇类三龄幼虫图解检索表

本文提供上海地区最常见嗜尸性蝇类三龄幼虫图解检索表,共4科11种.该检索表可用于案发现场法医昆虫的分类鉴定,进而推断尸体的死亡时间,也可用于重要有瓣蝇类幼虫的孽生地调查.     

特种五谷虫脂肪酸的体外抗肿瘤、抗HIV-1整合酶活性及组成分析

为研究特种五谷虫(大头金蝇Chrysomya megacephala蝇蛆的俗称)脂肪酸的体外抗肿瘤、抗HIV-1整合酶活性,并确定其组分构成,通过溶剂萃取和油脂酶解获得2种脂肪酸FA1和FA2,采用MTT法/SRB法测定其体外抗人白血病细胞HL-60/人肺癌细胞A-549活性,以及采用Biotin-ELISA法检测其对HIV-1整合酶的抑制作用;并由GC-MS分析确定其脂肪酸组成。结果表明：FA1和FA2对人白血病细胞/人肺癌细胞均有显著的抑制活性,其IC₅₀值在35～65 μg/mL;对HIV-1整合酶同样具有强烈的抑制活性,IC₅₀值分别为86.7 μg/mL和98.5 μg/mL。GC-MS分析表明,FA1和FA2化学组成相似,均含有15%～16%的多不饱和脂肪酸（PUFA),其中有2个组分为ω-6 PUFA。提示特定培养的五谷虫,其脂肪酸成分具有显著的体外抗肿瘤、抗HIV-1整合酶活性,其中含有的PUFA,尤其ω-6 PUFA,可能是主要活性组分;FA2在来源上和FA1存有相关性。     

药（毒）物对尸食性蝇类生长发育影响的研究进展

药（毒）物对尸食性蝇类生长发育影响是法医昆虫毒理学领域里一个十分重要的研究方向,其研究结果可对与药（毒）物相关死亡案件的死亡时间作出修正。随着近年来全球毒品及药物滥用情况的日趋严重,其所导致的死亡案件也越来越多。这类案件常常需要应用尸食性蝇类生长发育历期来推算死后经历时间（postmortem interval, PMI）。为了阐明该领域的研究进展以及未来研究的焦点和难点,本文在阐述法医昆虫毒理学概念和特点的基础上,按照不同的药（毒）物分类,对近年来药（毒）物对尸食性蝇类生长发育影响在国内外的研究进展进行了综述。研究表明,某些药（毒）物对尸食性蝇类生长发育具有一定的影响,且这种影响存在种属差异。目前,该领域的研究尚限于宏观现象观察阶段,其研究范围在不断拓宽,既有的研究也在进一步深化,但还有许多问题有待进一步探讨。     

Characterization of polymorphic microsatellite markers for the blowfly Chrysomya albiceps (Diptera: Calliphoridae)

Chrysomya albiceps is a blowfly of great medical, sanitary and forensic importance widely distributed in the Afrotropical, southern Palaearctic, northern Oriental regions and, recently, in Central and South Americas. Here, we report the characterization of 13 polymorphic microsatellite markers for C. albiceps. The number of alleles ranged from three to 13 alleles with expected heterozygosities ranging from 0.4668 to 0.8408. These markers will be extremely useful for investigating many important aspects of this species such as population structure, dispersal and colonization dynamics.     

吗啡对大头金蝇生长发育的影响及其用于推断PMI的初步探讨

用致死剂量盐酸吗啡注射家兔,处死后用其肌肉在恒温25℃条件下饲养大头金蝇Chrysomyia megacephala初孵幼虫,研究吗啡对大头金蝇生长发育的影响,及其用于法医学中推断死者死后间隔时间(Postmortem interval,PMI)方面的初步探讨.结果 显示,在25℃下孵化64 h后,处理组大头金蝇的体长、体重及蛹长、蛹重明显大于对照组幼虫(P＜0.05),这种趋势一直持续到羽化.吗啡缩短了大头金蝇的发育历期,根据大头金蝇发育历期进行PMI推断时,吗啡可使PMI推断值缩短14.7 h.     

The upper thermal tolerance for a Texas population of the hairy maggot blow fly Chrysomya rufifacies Macquart (Diptera: Calliphoridae)

1. The hairy maggot blow fly (Chrysomya rufifacies: Macquart) is an invasive necrophagous fly found throughout the continental United States. Chrysomya rufifacies is of medical/veterinary, forensic, and ecological importance due to its ability to cause myiasis, colonise human remains, and displace native Diptera. However, little is known about their upper thermal tolerance, which could be used to better predict their invasion potential.
2. We investigated the upper thermal tolerance of C. rufifacies exposed to different temperatures (20–45 °C), times (1–6 h), and nutrients (no food or water, water only, or a food-water mixture) for both sexes and two age ranges (young = 6–8 days post pupal emergence; old = 9–11 days post pupal emergence).
3. As temperature or duration increased, the probability of knockdown increased (0–100% at 20 and 45 °C and from 41 to 75% at 1 and 6 h), while the probability of survival decreased (99–2% at 20 and 45 °C and from 75 to 28% at 1 and 6 h). The availability of nutrients increased thermal tolerance at moderate temperatures (40 and 42 °C). Female flies were more thermally tolerant than males (probability of knockdown = 49% vs 58%; probability of survival = 58% vs 46%). Thermal tolerance did not differ by age.
4. These data reveal details about the upper thermal tolerance for a single population of C. rufifacies, and suggest that environmental and organismal factors ought to be considered in order to make meaningful predictions about the invasion potential of C. rufifacies in North America.

     

A simple, sterile food source for rearing the larvae of Lucilia sericata (Diptera: Calliphoridae)

arvae of the sheep blowfly Lucilia sericata (Meigen) were reared on various mixtures of puréed liver, agar, brewer's yeast and trypsin, in order to develop a simple, sterile, tissue-based diet. Growth and survival rates of larvae reared on a sterile 1:1 mixture of puréed liver with 3% Bacto agar equalled or exceeded those of larvae reared on raw liver. The addition of yeast and/or trypsin to the medium was of no additional benefit. This sterile, homogenous, tissue-based substrate offers a simple, convenient, inexpensive growth medium for rearing larvae for maggot therapy, and for testing the effects of various chemicals and dietary constituents on necrophagous insect larvae. It may be useful, therefore, in studies of myiasis, forensic entomology, and toxicology. This rearing medium also has the advantage that it can be stored for many months at room temperature without progressive decomposition or offensive odour.     

Effects of bait age, larval chemical cues and nutrient depletion on colonization by forensically important Calliphorid and Sarcophagid flies

Species colonization patterns on corpses and the frequency of carrion fly oviposition and larviposition are affected by decomposition stage and previous maggot colonization. This study investigated these effects on meat bait colonization by Victorian Diptera of forensic importance. Bait treatments were: 'aged' (aged for 4 days at 22 °C, allowing some decomposition); 'nutrient-depleted' [aged for 4 days at 22 °C with feeding Calliphora vicina (Robineau-Desvoidy) (Diptera: Calliphoridae) larvae]; 'extract' (fresh bait mixed with liquid formed by feeding C. vicina larvae), and 'fresh' (untreated control bait). Statistical analysis (α = 0.05) revealed that colonization frequency differed significantly among treatments (Welch's F(3,18.83) = 4.66, P < 0.05). Post hoc tests showed that fresh and extract baits were colonized extensively throughout the experiment with no significant difference, whereas the colonization of nutrient-depleted baits was significantly lower. This suggests that larval digestive enzymes, larval excreta and cuticular hydrocarbons have less effect on colonizing Diptera than the nutritional content of meat. The colonization of aged baits did not differ significantly from that of fresh, extract or nutrient-depleted baits. A further experiment testing 'very aged' (aged for 8 days at 28 °C), 'larvae-added' (fresh bait with C. vicina larvae added before placement) and 'fresh' (untreated control) baits revealed that very aged baits were colonized significantly less frequently than either fresh or larvae-added baits (Welch's F(2, 6.17) = 17.40, P < 0.05).